Specialized techniques for site-directed mutagenesis in cyanobacteria
... heterologous cassette (generally an antibiotic-resistance cassette) inserted within its open reading frame. However, as prokaryotic organisms have polycistronic genetic operons, the insertion of a selectable marker within an open reading frame can exert polar effects on downstream genes. Additionall ...
... heterologous cassette (generally an antibiotic-resistance cassette) inserted within its open reading frame. However, as prokaryotic organisms have polycistronic genetic operons, the insertion of a selectable marker within an open reading frame can exert polar effects on downstream genes. Additionall ...
New techniques and the GMO-legislation
... Techniques/methods of genetic modification yielding organisms to be excluded from the Directive, on the condition that they do not involve the use of recombinant nucleic acid molecules or genetically modified organisms other than those produced by one or more of the techniques/methods listed below a ...
... Techniques/methods of genetic modification yielding organisms to be excluded from the Directive, on the condition that they do not involve the use of recombinant nucleic acid molecules or genetically modified organisms other than those produced by one or more of the techniques/methods listed below a ...
An Escherichia coli Host Strain Useful for Efficient
... more or less similar (data not shown, but see Fig. 3). Several target gene constructs (including, for example, that encoding rat PTP-S [17]) which could be expressed in the latter strain only in the presence of plasmid pLysS (which encodes phage T7 lysozyme, an antagonist of T7 RNAP, and therefore s ...
... more or less similar (data not shown, but see Fig. 3). Several target gene constructs (including, for example, that encoding rat PTP-S [17]) which could be expressed in the latter strain only in the presence of plasmid pLysS (which encodes phage T7 lysozyme, an antagonist of T7 RNAP, and therefore s ...
pEGFP-N1 - ResearchGate
... Propagation in E. coli: • Suitable host strains: DH5a, HB101 and other general purpose strains. Single-stranded DNA production requires a host containing an F plasmid such as JM101 or XL1-Blue. • Selectable marker: plasmid confers resistance to kanamycin (30 µg/ml) to E. coli hosts. • E. coli replic ...
... Propagation in E. coli: • Suitable host strains: DH5a, HB101 and other general purpose strains. Single-stranded DNA production requires a host containing an F plasmid such as JM101 or XL1-Blue. • Selectable marker: plasmid confers resistance to kanamycin (30 µg/ml) to E. coli hosts. • E. coli replic ...
Recombinant DNA cloning technology
... a restriction map is often made to characterize the clone. A restriction map is a diagram showing where various restriction enzymes cut the DNA. If the sequence of the cloned fragment is known, the map can verify that the right fragment of DNA is cloned. If the sequence is not known, the map provide ...
... a restriction map is often made to characterize the clone. A restriction map is a diagram showing where various restriction enzymes cut the DNA. If the sequence of the cloned fragment is known, the map can verify that the right fragment of DNA is cloned. If the sequence is not known, the map provide ...
Temperate and lytic bacteriophages programmed to sensitize and
... the CRISPR array. Transcribed spacers guide Cas proteins to homologous sequences within the foreign nucleic acid, called protospacers, which are subsequently cleaved. The CRISPR-Cas systems have revolutionized molecular biology by providing efficient tools to precisely engineer genomes and manipulat ...
... the CRISPR array. Transcribed spacers guide Cas proteins to homologous sequences within the foreign nucleic acid, called protospacers, which are subsequently cleaved. The CRISPR-Cas systems have revolutionized molecular biology by providing efficient tools to precisely engineer genomes and manipulat ...
Differential effect of auxotrophies on the release of macromolecules
... S. typhimurium DthyA should not. Therefore, we equipped bacteria of both strains with an expression plasmid encoding firefly luciferase as a reporter. When both bacteria were transferred to medium without the complementing substrate, a rapid decrease in OD was observed for S. typhimurium Dasd, while ...
... S. typhimurium DthyA should not. Therefore, we equipped bacteria of both strains with an expression plasmid encoding firefly luciferase as a reporter. When both bacteria were transferred to medium without the complementing substrate, a rapid decrease in OD was observed for S. typhimurium Dasd, while ...
Chapter 13( Sample questions)
... Goals of genetic engineering include all of the following EXCEPT a. to learn more about genetic inheritance. b. to learn more about genetic diseases. c. to learn more about bacterial inheritance. d. to provide economic and social benefits. e. all of the above are goals of genetic engineering. Natura ...
... Goals of genetic engineering include all of the following EXCEPT a. to learn more about genetic inheritance. b. to learn more about genetic diseases. c. to learn more about bacterial inheritance. d. to provide economic and social benefits. e. all of the above are goals of genetic engineering. Natura ...
Design Genes with Ease Using In-Fusion® Cloning
... We observed fewer background colonies when the insertion site in a plasmid vector was created by digestion with two restriction enzymes rather than one. Treating the vector with phosphatase was unnecessary and reduced cloning efficiency. Accurate quantification and ratios of vector and DNA segments ...
... We observed fewer background colonies when the insertion site in a plasmid vector was created by digestion with two restriction enzymes rather than one. Treating the vector with phosphatase was unnecessary and reduced cloning efficiency. Accurate quantification and ratios of vector and DNA segments ...
DNA Technology
... transformation (recall from chapter 10) • A cell takes in DNA from outside the cell and becomes a part of that organism’s genome ...
... transformation (recall from chapter 10) • A cell takes in DNA from outside the cell and becomes a part of that organism’s genome ...
Experiments Covered by the NIH Guidelines
... Lambda or lambdoid bacteriophages or Ff non-conjugative plasmids are used as vectors (unless the DNA inserted into E. coli K-12 is from a prokaryote that naturally exchanges genetic information with E. coli, in which case any E. coli K-12 vector may be used). ...
... Lambda or lambdoid bacteriophages or Ff non-conjugative plasmids are used as vectors (unless the DNA inserted into E. coli K-12 is from a prokaryote that naturally exchanges genetic information with E. coli, in which case any E. coli K-12 vector may be used). ...
Section 13-1 Ghanging the Living World
... would like an animal to have. Thebreeder then chooses a male and female animal that have those traits and breeds them. The breeder expects that they will have offspring with the same traits. lfu.!:tfu tle yaryts l brceder would sclect in order to breeil offspring ...
... would like an animal to have. Thebreeder then chooses a male and female animal that have those traits and breeds them. The breeder expects that they will have offspring with the same traits. lfu.!:tfu tle yaryts l brceder would sclect in order to breeil offspring ...
AP Biology
... spontaneous mutations for 1 gene, only ~1 mutation in 10 million replications each day, ~2,000 bacteria develop mutation in that ...
... spontaneous mutations for 1 gene, only ~1 mutation in 10 million replications each day, ~2,000 bacteria develop mutation in that ...
Vectors and Libraries
... vector backbone and transformed into bacteria. Each individual bacterial transformant (clone) will contain a different fragment of the genome. All of the plasmids in a particular cell will contain the same insert. As the plasmid replicates there will be 50-100 copies of the plasmid in each cell. In ...
... vector backbone and transformed into bacteria. Each individual bacterial transformant (clone) will contain a different fragment of the genome. All of the plasmids in a particular cell will contain the same insert. As the plasmid replicates there will be 50-100 copies of the plasmid in each cell. In ...
C2005/F2401 `09
... B-3. The transformed cells would NOT make any toxin if the plasmid contained a deletion of (gene 1) (gene 2) (gene 3) (gene 4) (gene 5) (gene 6) (P2) (none of these – cells would make some toxin no matter what). B-4. These cells would make LOW levels of toxin (<10% of normal) if the plasmid containe ...
... B-3. The transformed cells would NOT make any toxin if the plasmid contained a deletion of (gene 1) (gene 2) (gene 3) (gene 4) (gene 5) (gene 6) (P2) (none of these – cells would make some toxin no matter what). B-4. These cells would make LOW levels of toxin (<10% of normal) if the plasmid containe ...
Ch. 1 Plasmids
... vector backbone and transformed into bacteria. Each individual bacterial transformant (clone) will contain a different fragment of the genome. All of the plasmids in a particular cell will contain the same insert. As the plasmid replicates there will be 50-100 copies of the plasmid in each cell. In ...
... vector backbone and transformed into bacteria. Each individual bacterial transformant (clone) will contain a different fragment of the genome. All of the plasmids in a particular cell will contain the same insert. As the plasmid replicates there will be 50-100 copies of the plasmid in each cell. In ...
Chaperone Competent Cell BL21
... The chaperone plasmids carry a pACYC origin of replication plus a chloramphenicol resistance which allows their use with most common ColE1-type plasmids having ampicilin resistance gene as a marker. As each chaperone gene is located at the downstream of araB or Pzt-1 (tet) promoter, separate express ...
... The chaperone plasmids carry a pACYC origin of replication plus a chloramphenicol resistance which allows their use with most common ColE1-type plasmids having ampicilin resistance gene as a marker. As each chaperone gene is located at the downstream of araB or Pzt-1 (tet) promoter, separate express ...
CopyRight® v2.0 Fosmid Cloning Kit
... High DNA yields.The CopyRight v2.0 Fosmid vector and Replicator FOS cells feature inducible amplification of copy number**, increasing yields to as many as 50 copies per cell. CopyRight amplification is more robust than the similar CopyControl system (Figure 4) and permits easy isolation of plasmid ...
... High DNA yields.The CopyRight v2.0 Fosmid vector and Replicator FOS cells feature inducible amplification of copy number**, increasing yields to as many as 50 copies per cell. CopyRight amplification is more robust than the similar CopyControl system (Figure 4) and permits easy isolation of plasmid ...
Control of Gene Expression
... 4. use representations to describe how gene regulation influences cell products and function. Gene expression is controlled by environmental signals and developmental cascades that involve both regulatory and structural genes. A variety of different gene regulatory systems are found in nature. Two o ...
... 4. use representations to describe how gene regulation influences cell products and function. Gene expression is controlled by environmental signals and developmental cascades that involve both regulatory and structural genes. A variety of different gene regulatory systems are found in nature. Two o ...
Document
... • Genetic engineering involves changing an organism’s DNA to give it new traits by inserting cloned genes from one organism into a different organism. – Possible because the genetic code is shared by all organisms (all living things share the same 4 nucleotides A,T,C,G) ...
... • Genetic engineering involves changing an organism’s DNA to give it new traits by inserting cloned genes from one organism into a different organism. – Possible because the genetic code is shared by all organisms (all living things share the same 4 nucleotides A,T,C,G) ...
... and 2% Triton X-100 was then applied to the replica print. After incu/j-oxidation bation at 37 "C for 1 h, thereplica print was washed twicein chloroform: FIG.1. Pathways for the incorporation of exogenous fatty ac- methano1:acetic acid (3:6:1, v/v) to remove unreacted [3H]palmitic acid ids into pho ...
Biotechnology toolkit part 2
... piece of DNA of interest can be cut with the same restriction enzyme and inserted into the plasmid. This can then be put back into the bacteria. The bacterium divides; every time it does it replicates the foreign DNA until you have many copies. Plasmids are useful because They can be taken up by b ...
... piece of DNA of interest can be cut with the same restriction enzyme and inserted into the plasmid. This can then be put back into the bacteria. The bacterium divides; every time it does it replicates the foreign DNA until you have many copies. Plasmids are useful because They can be taken up by b ...
Insertion of liver enriched transcription
... regulatory system. Tissue-specific promoters or enhancers are in use in transgenic animals and could be utilized in medicine for gene therapy. At present the usual method for selection of a tissue-specific promoter is to identify a gene, which is expressed at unusually high level in the target tissu ...
... regulatory system. Tissue-specific promoters or enhancers are in use in transgenic animals and could be utilized in medicine for gene therapy. At present the usual method for selection of a tissue-specific promoter is to identify a gene, which is expressed at unusually high level in the target tissu ...
Plasmid
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A plasmid is a small DNA molecule within a cell that is physically separated from a chromosomal DNA and can replicate independently. They are most commonly found in bacteria as small, circular, double-stranded DNA molecules; however, plasmids are sometimes present in archaea and eukaryotic organisms. In nature, plasmids often carry genes that may benefit the survival of the organism, for example antibiotic resistance. While the chromosomes are big and contain all the essential information for living, plasmids usually are very small and contain only additional information. Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms.Plasmids are considered replicons, a unit of DNA capable of replicating autonomously within a suitable host. However, plasmids, like viruses, are not generally classified as life. Plasmids can be transmitted from one bacterium to another (even of another species) via three main mechanisms: transformation, transduction, and conjugation. This host-to-host transfer of genetic material is called horizontal gene transfer, and plasmids can be considered part of the mobilome. Unlike viruses (which encase their genetic material in a protective protein coat called a capsid), plasmids are ""naked"" DNA and do not encode genes necessary to encase the genetic material for transfer to a new host. However, some classes of plasmids encode the conjugative ""sex"" pilus necessary for their own transfer. The size of the plasmid varies from 1 to over 200 kbp, and the number of identical plasmids in a single cell can range anywhere from one to thousands under some circumstances.The relationship between microbes and plasmid DNA is neither parasitic nor mutualistic, because each implies the presence of an independent species living in a detrimental or commensal state with the host organism. Rather, plasmids provide a mechanism for horizontal gene transfer within a population of microbes and typically provide a selective advantage under a given environmental state. Plasmids may carry genes that provide resistance to naturally occurring antibiotics in a competitive environmental niche, or the proteins produced may act as toxins under similar circumstances, or allow the organism to utilize particular organic compounds that would be advantageous when nutrients are scarce.